KR101905865B1 - Composition Comprising Sarpogrelate for Preventing or Treating Sensorineural Hearing Loss - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for preventing, ameliorating or treating sensorineural hearing loss containing a sarpogrelate or a pharmaceutically acceptable salt thereof as an active ingredient. The sarpogrelate according to the present invention can be effectively used for preventing and ameliorating sensorineural hearing loss by protecting hearing from the noise by suppressing auditory cell death and increasing expression of antioxidant enzymes in auditory cells.

Description

TECHNICAL FIELD [0001] The present invention relates to a composition for preventing or treating sensory neural deafness,

The present invention relates to a composition for preventing, ameliorating or treating sensory neural deafness containing as an active ingredient a saporiglate or a pharmaceutically acceptable salt thereof.

Hearing loss, or hearing loss, is a very common disorder in about 15-20% of the population. Due to the environmental pollution and aging of modern society, hearing impairment is increasing and the hearing impairment is permanent, so it is very important to prevent it before it occurs. The occurrence of hearing loss is mostly caused by environmental factors and genetic factors such as suddenness, weak physical properties (antibiotics such as aminoglycosides, anticancer drugs), noise, trauma, senility and congenital, It is caused by sensory nerve impairment characterized by death.

There are many signaling pathways involved in the regeneration of inner ear hair cells, the proliferation and differentiation of hair cells, and the use of gene therapy or cell transplantation (Hanyang Med Rev, 2005). However, there are few studies on the mechanism of prevention and prevention of hearing loss and the development of preventive and therapeutic agents (Hanyang Med Rev, 2015).

As the society has become industrialized, the number of hearing impairments due to noise has also increased rapidly in recent years. In addition to occupational noise-induced hearing loss of workers working in noisy environments, noise-induced hearing loss due to cultural and leisure activities is also increasing. Human auditory organs have been reported to be affected by noise above 75 dBA, and noise of 75 dBA is the noise level of the road on which the car is traveling. It is believed that in industrial society, everyone lives in a level of harmful noise to auditory organ have. In addition to environmental noise, which is inevitably heard, many people are exposed to loud sounds even in leisure activities including MP3 use. Recently, noise-induced hearing loss has appeared in various ages. When young, noise-induced hearing loss becomes more and more severe when he is accompanied by aging. That is, when the younger generation experiencing noise-induced hearing loss becomes an elderly person, the degree of hearing loss becomes worse, and hearing loss has an important influence on the quality of life of various generations, from the elderly to the young.

Antioxidant therapy with antioxidants has been the main research aimed at the prevention and treatment of noise-induced hearing loss. Among them, vitamin E, aspirin and N-acetylcysteine have been shown to reduce toxicity by aminoglycoside antibiotics , But there is no clear prophylactic agent yet. Despite many attempts to prevent and treat hearing loss, clear molecular mechanisms of hearing loss and preventive and therapeutic measures have yet to be presented. to be.

Sarpogrelate is a platelet aggregation inhibitor with a new mechanism of action that selectively antagonizes the serotonin receptor (5-HT2) in platelets and blood vessels, one of several agonists that activate platelets. It is known that it blocks the metabolism of 5-HT which causes thrombus formation and vascular occlusion by stimulating vasoconstriction and smooth muscle cell increase while being stored and secreted in platelet. In addition, it is known that hearing loss occurs due to microvascular disturbance and vasoconstriction in various causes of hearing loss, which deteriorates auditory and auditory cell functions.

Accordingly, the present inventors have made intensive efforts to find out the molecular mechanism of hearing loss induced by noise and to prevent and treat hearing loss. As a result, it has been found that the sapoglirate improving microvascular disorder improves hearing loss and hearing in an animal model of noise- And the present invention has been completed.

It is an object of the present invention to provide a composition for treating sensory neural deafness, which comprises sarpogrelate or a pharmaceutically acceptable salt thereof as an active ingredient, a composition for treating said sensory neural deafness, a composition comprising said ginseng extract, ginkgo leaf extract, resveratrol, Wherein the active ingredient is at least one selected from the group consisting of vitis vinifera leaf dry extract and sarpogrelate or a pharmaceutically acceptable salt thereof, The present invention provides a composition for co-administration with at least one selected from the group consisting of Korean red ginseng extract, ginkgo biloba extract, resveratrol, and vitis vinifera leaf dry extract.

In order to achieve the above object, the present invention provides a pharmaceutical composition for preventing or treating sensory neural deafness, comprising sarpogrelate or a pharmaceutically acceptable salt thereof as an active ingredient.

The present invention also relates to a pharmaceutical composition comprising sarpogrelate or a pharmaceutically acceptable salt thereof and a ginseng ginseng extract, a ginkgo leaf extract, resveratrol, vitis vinifera leaf dry extract The present invention provides a combined preparation for the treatment of sensory neural deafness, comprising at least one selected from the group consisting of

The present invention also relates to a method for the treatment and / or prophylaxis of ginseng extract, ginkgo leaf extract, resveratrol, and vitis vinifera leaf extract containing sarpogrelate or a pharmaceutically acceptable salt thereof as an active ingredient dry extract of the present invention.

The present invention also provides a food composition for preventing or ameliorating sensory neural deafness containing sarpogrelate or a pharmaceutically acceptable salt thereof as an active ingredient.

The composition comprising sarpogrelate according to the present invention protects the hearing caused by noise by suppressing auditory cell death and increasing the expression of antioxidant enzymes in auditory cells, Can be usefully used.

Figure 1 shows a schematic diagram of an in-vivo experiment using a Balb / c mouse.
FIG. 2 shows the result of measurement of the left and right ABR (auditory brainstem response) after the pre-treatment with the sampogrirate and before the exposure, the 1 day, 1 week, and 2 weeks after the exposure.
Fig. 3 shows the expression of antioxidants (catalase: green, SOD2: red, DAPI: blue) by the sapoglylate in the spiral ganglia (apex, middle and base) of the mouse with noise-induced hearing loss.
FIG. 4 shows the expression of antioxidant substances (catalase: green, SOD2: red, DAPI: blue) caused by sapoglylate in auditory hair cells in the woah of a mouse with noise-induced hearing loss.
Figure 5 after sanding draw rate pretreated hearing cell line, HEI-OC1 cell, H ₂ O ₂ And the inhibition of cytotoxicity and death by treatment was confirmed by WST-1 analysis.
FIG. 6 shows mRNA expression of antioxidants (catalase and SOD2) in auditory cells treated with H ₂ O ₂ and sapoglylate through quantitative real-time PCR.
FIG. 7 is a western blot image showing protein expression of antioxidants (catalase and SOD2) in auditory cells treated with H ₂ O ₂ and saponylate.

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In general, the nomenclature used herein is well known and commonly used in the art.

In the present invention, it was confirmed that hearing loss reduction was suppressed and improved by inhibiting apoptosis and inducing antioxidant enzymes in cells by treating sarpogrelate after establishing a model of noise-null hearing loss mouse and a noise-canceling hearing cell line . In other words, the prevention, amelioration and therapeutic effect of the sensory neural deafness due to the sapoglylate was confirmed.

Accordingly, the present invention relates to a pharmaceutical composition for preventing or treating sensory neural deafness, which comprises, as an active ingredient, sarpogrelate or a pharmaceutically acceptable salt thereof in one aspect.

The sarpogrelate of the present invention is represented by the following Formula 1 and the IUPAC name is 4- [2- (dimethylamino) -1- ({2- [2- (3-methoxyphenyl) ethyl] phenoxy } Methyl) ethoxy] -4-oxobutanoic acid.

[Chemical Formula 1]

The sampoglirate exhibits antagonistic action on serotonin receptors of platelets and vascular smooth muscle, thus exhibiting anti-platelet action and vasoconstricting action. It is possible to confirm platelet aggregation inhibition by simultaneous addition of serotonin and collagen in healthy adults and patients with chronic arterial occlusion, inhibit platelet aggregation by collagen and secondary aggregation of platelets by ADP or epinephrine, Platelet aggregation is enhanced by serotonin, which inhibits enhanced platelet aggregation. In addition, the sapogenirate suppresses the development of the pathology in the peripheral arterial occlusion model (rat peripheral artery embolization by lauric acid injection), and the arterial blood pressure model (the arterial blood supply due to vascular endothelial damage, the arterial blood supply to the polyethylene tube- Thrombotic effects that inhibit the formation of thrombus. In vitro experiments using rat vascular smooth muscle have shown that sapogenate suppresses the contraction of vascular smooth muscle caused by serotonin and inhibits vasoconstriction by inhibiting the contraction when vascular smooth muscle contracts with platelet aggregation.

Therefore, the sapoglylate is commercially available as a hydrochloride salt for the improvement of ischemic symptoms such as ulcer, pain and cold feeling caused by chronic arterial occlusion (Burger's disease, occlusive atherosclerosis, diabetic peripheral vasculopathy, etc.). However, to date, no document has been reported to show a direct association of sampogirate with hearing loss.

In the present invention, the composition containing sarpogrelate is selected from the group consisting of Korean red ginseng extract, ginkgo biloba extract, resveratrol, and vitis vinifera leaf dry extract. But the present invention is not limited thereto.

The Korean red ginseng refers to ginseng produced in Korea and is representative of medicinal plants. The term " Korean red ginseng " Means Korean red ginseng obtained from the society, but not limited thereto. Also, Korean red ginseng includes all of those classified as fresh ginseng, red ginseng, and white ginseng. It is said that the original shape of Korean red ginseng remains intact, and the processed ginseng, which is processed without processing the original shape, is said to be processed products.

The classification of Korean red ginseng differs depending on the manufacturing method and appearance. The red ginseng and dried ginseng are classified as red ginseng and white ginseng products, respectively. Consideration of invention Red ginseng includes, but is not limited to all of these.

The red ginseng extract of the present invention can be extracted using an organic solvent such as water or alcohol as an extraction solvent. Specifically, there may be mentioned water, a functional or anhydrous lower alcohol having 1 to 4 carbon atoms, a mixed solvent of the lower alcohol and water, or an extraction solvent such as acetone, ethyl acetate, chloroform, 1,3-butylene glycol, ≪ / RTI > Preferably, the red ginseng extract can be prepared using a functional lower alcohol, most preferably ethanol. The extract of the present invention includes not only the above-mentioned extraction solvent but also red ginseng extract which has substantially the same effect as that produced by using other extraction solvent.

In addition, the red ginseng extract of the present invention includes not only the extract obtained by the above-mentioned extraction solvent, but also the extract obtained through a conventional purification process. For example, separation using an ultrafiltration membrane having a constant molecular weight cut-off value, separation obtained by various chromatography (size, charge, prepared by separation for hydrophobicity or affinity) The fraction is also included in the red ginseng extract of the present invention. In addition, the red ginseng extract of the present invention can be prepared in powder form by an additional process such as vacuum distillation and freeze-drying or spray drying.

The 'Ginkgo biloba extract (GbE)' of the present invention is preferably EGb761, but is not limited thereto. In addition, the bank side extract is composed of 24% of flavonoid and 6% of terpenoid, and is known to be effective for bronchial asthma, bronchitis, peripheral blood disorder, and brain dysfunction.

Resveratrol (3, 5, 4-trans-trihydroxy stilbene, Resveratrol) of the present invention belongs to the polyphenol family of antioxidants found in plants, And is known to be effective in preventing blood clotting, a dangerous phenomenon that causes heart attacks and strokes. The 'vitis vinifera leaf dry extract' of the present invention is also known to contain a large amount of antioxidants. Resveratrol is more effective than vitamin C in clearing hydroxyl radicals, and adding resveratrol to vitamin A can have an additive effect. The use of resveratrol in combination with other antioxidants (not vitamins A, C or E, not vasodilator magnesium or other vasodilator substances) is known to reduce aging-related hearing loss.

Among the various causes of hearing loss, smoking and drinking are major deteriorating factors of chronic diseases, leading to microvascular disturbance, which is likely to result in hearing loss, and stresses that cause vasoconstriction may also impair auditory function of auditory and auditory cells. It is known to give. In other words, since hearing loss may occur due to microvascular disturbance and vasoconstriction, various approaches to the hearing loss induction mechanism using the inhibitor of platelet aggregation, such as sapoglylate, and the development of a medicament for preventing noise-induced hearing loss, It is expected to maximize the effect of treatment for hearing loss.

In order to overcome the period of clinical trial development and the period of development of new drugs which have not developed definitive treatment yet, hearing loss has been developed by using new drug re- The grill rate can be applied to the prevention and treatment of noise-induced hearing loss. In addition, by applying the action mechanism of the sampogirate, it is possible to develop other preventive and therapeutic agents for toxic, hearing loss, and agoraphobic hearing loss, and it can be used as a combination therapy with other treatment methods for hearing loss patients. This is also possible to prevent the hearing loss caused by other causes and to develop the therapeutic drug as zero.

The term " sensorineural hearing loss "of the present invention occurs when the component of the inner ear or the associated nerve component is affected, and when the auditory or auditory pathway of the brain is affected, ≪ / RTI > Sensory hearing loss can be hereditary, or it can be due to acoustic trauma (eg, very loud noises such as explosions), viral infections, drug-induced or Meniere's disease. Neurotic deafness can be caused by a brain tumor, infection or various brain and neurological disorders, such as stroke. Some genetic disorders, such as Levofloxacin (a defective accumulation of branched fatty acids), can also cause neurological disorders that affect hearing loss. The auditory nerve pathway may be used for the treatment of dehydrative diseases such as idiopathic inflammatory dehydratative diseases (including multiple sclerosis), transverse myelitis, Deby's disease, progressive multiseptic leukopenia, Guillain-Barré syndrome, chronic inflammatory dehydrative polyneuropathy, MAG is impaired by peripheral neuropathy.

In the present invention, the sensory neural deafness is preferably, but not limited to, a noise-induced hearing loss, a toxic hearing loss, a sudden hearing loss or an aging hearing loss. In addition, the sensory neuron deafness may be characterized by damage to inner ear hair cells and surrounding tissues.

The " noise-induced hearing loss " of the present invention can lead to hearing loss even when exposed to loud noises such as loud music, heavy equipment or mechanical devices, airplanes, bombardment or other human noise for a long period of time. Deafness is caused by the destruction of the inner ear's hair cell receptors. Such hearing loss often involves tinnitus. Often, permanent hearing loss is diagnosed. Currently, there are no treatments for noise-induced hearing loss, but several therapies have been experimentally developed, including treatment with insulin-like growth factor 1 (IGF-1) (Lee et al. Otol . Neurotol. 28: 976-981, 2007 ).

The "aging-induced hearing loss " or " aging-induced hearing loss" of the present invention occurs as a part of normal aging due to age-related hearing loss, and is caused by denaturation of receptor cells in the inner ear cortisone organs. Another cause may be the loss of flexibility of the basilar membrane of the womb, as well as the decrease in the number of nerve fibers in the vestibular nerve. There is currently no known cure for permanent hearing loss due to senile hearing loss or excessive noise.

In the present invention, the composition may be characterized by suppressing apoptosis of auditory cells or increasing expression of antioxidant enzymes in auditory cells.

The term " wow " of the present invention is an inner ear part related to hearing. The wah is a spiral tube-like structure wrapped in a snail-like shape. The inside of the wah is divided into three areas, and is divided by the location of the vestibular and basilar membrane. The area above the vestibule is the anterior chamber, which contains the outer lymph fluid, which extends from the innermost to the apex of the wah and has a low potassium content and a high sodium content. The basement membrane confines the hyperreal area, which extends from the vertex of the wah to the gill window and also contains the outer lymph fluid. The basement membrane contains a large number of stiff fibers, which gradually increase in length from the window to the apex of the wah. The fibers of the basement membrane are vibrated when activated by sound. The cochlea is located between the anterior and the posterior, and the distal end of the cochlea is closed at the apex of the wah. The cochlea contains the lymphatic fluid, which is similar to cerebrospinal fluid and has a high potassium content.

The auditory organ, the "cortical organ," is located on the basement membrane and extends upwardly into the cochlea. The Corti organ contains hair cells with hairy protrusions extending from the free surface and contacts a gelatin surface called the covering membrane. Although the hair cells do not have axons, they are surrounded by sensory nerve fibers that form the womb of the vestibular nerve (cranial nerve VIII).

In another aspect, the present invention relates to a pharmaceutical composition comprising sarpogrelate or a pharmaceutically acceptable salt thereof and a red ginseng extract, a ginkgo extract, a resveratrol and a vitis vinifera leaf dry extract, The present invention relates to a combination preparation for preventing or treating sensory neural deafness.

In the present invention, the combined preparation is preferably a tablet, a foamable tablet, a granule, a powder, an injection, a pill or a capsule, but is not limited thereto.

In formulating the two active ingredients into a single formulation, the physico-chemical properties of the two active ingredients and the impact on bioavailability and stability due to in vivo / in vitro interaction between the two active ingredients should be considered important .

In the present invention, the pharmaceutically acceptable salts may include both non-toxic inorganic acid salts and organic acid salts, for example, hydrochloride, sulfate, mesylate, malate, maleate, mesylate, But is preferably at least one salt selected from the group consisting of oxalate, fumarate, tartarate, citrate, succinate, acetate and phosphate, with hydrochloride being most preferred, but not limited thereto.

Said sapoglylate or a pharmaceutically acceptable salt thereof is commercially available or can be easily prepared by methods known in the art (European Patent Publication No. 0072942)

In other embodiments of the present invention, suitable carriers, excipients, disintegrants, sweeteners, coatings, swelling agents, lubricants, lubricants, lubricants, and the like, which are conventionally used in the preparation of compositions comprising sanguoglate or a pharmaceutically acceptable salt thereof, And may further comprise at least one additive selected from the group consisting of flavor agents, antioxidants, buffers, bacteriostats, diluents, dispersants, surfactants, binders and lubricants.

Specific examples of carriers, excipients and diluents include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, Cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil. Solid formulations for oral administration may be in the form of tablets, pills, powders, granules, capsules These solid preparations can be prepared by mixing at least one excipient, for example, starch, calcium carbonate, sucrose or lactose, gelatin, etc., into the composition. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, syrups and the like, and various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included in addition to water and liquid paraffin which are commonly used simple diluents. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, suppositories, and the like. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. As the suppository base, witepsol, macrogol, tween 61, cacao paper, laurin, glycerogelatin and the like can be used.

The pharmaceutical composition according to the present invention may be administered orally (e.g., by taking or inhalation) or parenterally (e.g., by injection, sedation, implantation, suppository), and the injection may be, for example, , Subcutaneous injection, intramuscular injection or intraperitoneal injection. The pharmaceutical composition according to the present invention may be formulated into tablets, capsules, granules, fine subtilae, powders, sublingual tablets, suppositories, ointments, injections, emulsions, suspensions, syrups, Can be formulated. The various forms of the pharmaceutical composition according to the present invention can be prepared by a known technique using a pharmaceutically acceptable carrier commonly used in each formulation. Examples of pharmaceutically acceptable carriers are excipients such as excipients, binders, disintegrating agents, lubricants, preservatives, antioxidants, isotonic agents, buffers, encapsulating agents, sweeteners, solubilizers, bases, , Suspending agents, stabilizers, coloring agents and the like.

The pharmaceutical composition according to the present invention varies depending on the form of the medicament, but includes about 0.01 to 95% by weight of the compound of the present invention (saporoglate).

The amount of the saponylate used as an active ingredient of the pharmaceutical composition according to the present invention may vary depending on the patient's age, sex, weight, and disease, but is preferably 0.001 to 100 mg / kg, preferably 0.01 to 10 mg / Or several times.

In addition, the dose of the sapogrylate according to the present invention may be increased or decreased depending on the route of administration, degree of disease, sex, weight, age, and the like. Thus, the dosage amounts are not intended to limit the scope of the invention in any manner.

The pharmaceutical composition may be administered to mammals such as rats, mice, livestock, humans, and the like in a variety of routes. All modes of administration may be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intratracheal, intrauterine or intracerebroventricular injections.

The term "administering" as used herein means introducing the composition of the present invention to a patient in any suitable manner, and the administration route of the composition of the present invention may be administered through any conventional route so long as it can reach the target tissue have.

The present invention, in another aspect, relates to a method for the treatment and / or prevention of a Korean red ginseng extract, a ginkgo leaf extract, a resveratrol and a European grape-side dried extract (hereinafter referred to as " ginseng extract ") containing sarpogrelate or a pharmaceutically acceptable salt thereof as an active ingredient vitis vinifera leaf dry extract). < / RTI >

In the present invention, it is preferable to administer each of the existing single preparations in combination, and the above sapoglirate may be added to the Korean red ginseng extract, the bank leaf extract, the resveratrol and the vitis vinifera leaf dry extract And one or more selected from the group consisting of the above-mentioned compounds.

In the present invention, the combined administration is preferably performed sequentially, simultaneously or in reverse order, but is not limited thereto.

The term "pharmaceutically effective amount" of the present invention means that the composition of the present invention can be administered in a pharmaceutically effective amount, which is sufficient to treat or prevent the disease at a reasonable benefit / risk ratio applicable to medical treatment or prevention And the effective dose level is determined according to the severity of the disease, the activity of the drug, the age, body weight, health, sex, sensitivity of the patient to the drug, administration time of the composition of the present invention, , Factors including drugs used in combination with or co-used with the compositions of the present invention used, and other factors well known in the medical arts. The pharmaceutical composition of the present invention may be administered as an individual therapeutic agent or in combination with another therapeutic agent, and may be administered sequentially or simultaneously with a conventional therapeutic agent. And can be administered singly or multiply. It is important to take into account all of the above factors and administer an amount that will achieve the maximum effect in the least amount without side effects.

In another aspect, the present invention relates to a food composition for preventing or ameliorating sensory neural deafness containing sarpogrelate or a pharmaceutically acceptable salt thereof as an active ingredient.

In the present invention, it is preferable to further include at least one selected from the group consisting of Korean red ginseng extract, ginkgo biloba extract, resveratrol and vitis vinifera leaf dry extract, It is not.

In the present invention, the sensory neural deafness may be characterized by being a noise-induced hearing loss, a toxic hearing loss, a sudden hearing loss or an aging hearing loss, And may be characterized by damage to the tissue.

In the present invention, it is preferable, but not limited, to further include pharmaceutically acceptable additives.

The food composition may be provided in the form of powder, granules, tablets, capsules, syrups or beverages. The health food may be used in combination with other food or food additives in addition to the active ingredient, sapoglylate, Can be used. The amount of the active ingredient to be mixed can be suitably determined according to its use purpose, for example, prevention, health or therapeutic treatment.

The effective dose of the sampoglirate contained in the food composition may be used in accordance with the effective dose of the pharmaceutical composition but may be less than the above range for health and hygiene purposes or for long term consumption for health control purposes And it is clear that the active ingredient can be used in an amount exceeding the above range since there is no problem in terms of safety.

There is no particular limitation on the kind of the food composition. Examples of the food composition include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, , A drink, an alcoholic beverage, and a vitamin complex, all of which include health foods in a conventional sense.

The food composition may comprise a health beverage composition, and may contain various flavors or natural carbohydrates, such as ordinary beverages, as an additional ingredient. The natural carbohydrates are sugar monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol. Examples of sweeteners include natural sweeteners such as tau Martin and stevia extract, synthetic sweeteners such as saccharin and aspartame, and the like.

[ Example ]

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are only for illustrating the present invention and that the scope of the present invention is not construed as being limited by these embodiments.

Example  1: Using an animal model of noise-induced hearing loss Invivo  (in- vivo ) Experiment

1-1: Establishment of animal model for noise-induced hearing loss

To exclude mice with congenital abnormal hearing, prior hearing tests were performed on all mice before the experiment. A total of 24 male and 7 week old Balb / c mice underwent auditory brainstem response (ABR) test. Twelve experimental groups were administered sopoggylate (50 mg / kg) And 12 control groups without administration.

(50 mg / kg) was administered once a day for 7 days, and an animal model in which noise-induced hearing loss was induced by establishing a white noise of 100 dB for 50 minutes was established (FIG. 1) .

1-2: On the sapphire grill rate  Hearing protection by

Auditory brainstem response (ABR test) was performed 7 days after the administration of saporiglirate (50 mg / kg) to analyze auditory function.

TDT (Tucker-Davis Technologies) Using a ABR device, give a tone burst of 8 kHz, 16 kHz, and 32 kHz to determine the minimum stimulus volume (dB) at which the waveform of wave V (wave V) The audiograms of the control group and the control group, which were administered with the sampogirate drug before the noise exposure and the control group without the drug, were compared and analyzed to prevent the noise - induced hearing loss of the sampogirate.

As a result, hearing thresholds were observed to increase at 1 day, 7 days, and 14 days after noise exposure. However, it was confirmed that the hearing threshold of the experimental group pretreated with the sampogirrile was significantly lower than that of the control group (Fig. 2). That is, it has been found that the sampogirate prevents and protects against hearing loss due to noise.

Example  2: Tissue Dyeing Analysis of Animal Models with Noise

Scanning electron microscope (SEM) and immunohistochemistry were performed for morphological analysis of helical spinal ganglia and Corti auditory hair cells of the animal model induced by the noise-induced hearing loss of Example 1.

The expression of antioxidant - related proteins was observed by immunohistochemical staining in the spiral ganglia and auditory hair cells of the apex, middle and base of the experimental group and the control group pretreated with the sampogirate. After the animal tissues were fixed, the sections were embedded in paraffin to prepare tissue sections. Antioxidant-related proteins were firstly attached to the cells and secondary antibody was attached to the cells. Immunohistochemistry was performed to analyze the expression pattern of the antioxidant-related proteins. Antibodies were stained with antioxidant catalase (Abcam, ab1877, Cambridge, UK) and SOD2 (Santa Cruz, CA, USA) and DAPI (Invitrogen, Carlsbad, CA).

As a result, it was found that the expression of antioxidant enzymes catalase and SOD2 was significantly increased in the experimental group pretreated with saporiglate (Figs. 3 and 4). That is, it was found that the sampoglirate protects the hearing loss caused by the noise.

Example  3: On the sapphire grill rate  by Auditory cell  Cell death suppression

A; (House-Ear Institute-organ of Corti 1 HEI-OC1) a was used, 33 ℃, 10% CO ₂ high concentration is added a DMEM medium (Dulbecco's modified Eagle's Medium) glutamine in terms hearing cell line expressing the hearing gene 10% fetal bovine serum (FBS) and 25 U / ml interferon gamma were added and HEI-OC1 cells were cultured.

To induce oxidative stress - induced oxygen species (ROS) in the process of hearing loss induced by noise, H ₂ O ₂ was applied to establish a noise - canceling cell line.

After 24, 24 hours of treatment with the sampogirate 0.1, 1, 10 and 25 μM, the cytotoxicity of the sampogrilate was measured by WST-1 assay. Further, quantitatively H ₂ over the sandpaper draw rates of 0.1, 1, 10 and after pre-treatment for the 25 μM 1 sigan, WST-1 (Takara-Bio , Tokyo, Japan) with H ₂ O ₂ 500μM were treated for 24 hours Analysis The cytotoxicity and apoptotic effects of O ₂ were evaluated.

As a result, there was no cytotoxicity due to the sapoglyreate, and the sapoglirate showed the effect of inhibiting H ₂ O _2- induced apoptosis of auditory cells (FIG. 5).

Example  4: On the sapphire grill rate  by Auditory cell  Antioxidant effect

The antioxidant activity by saponylate in the noise-induced hearing loss cell line by the H ₂ O ₂ treatment of Example 3 was analyzed.

After the pretreatment of 1, 10, 25 and 50 μM of the saporoglirate for 1 hour, 250 μM of H ₂ O ₂ was treated for 24 hours, and mRNA and protein expression of the antioxidant enzymes catalase and SOD2 (superoxide dismutase 2) were confirmed . mRNA expression was quantified by quantitative real-time PCR after total RNA was extracted and cDNA was synthesized. Protein expression was measured by SDS-PAGE followed by catalase (Abcam, ab 1877, Cambridge, UK) and SOD2 (Superoxide dismutase 2, Santa Cruz, CA, USA) Antibodies were attached to confirm protein expression.

As a result, it was confirmed that the expression of mRNA and protein of catalase and SOD2 was increased in the group treated with the saporogylate (FIGS. 6 and 7). In other words, the in-vitro results using the auditory cell line also showed the effect of the hearing protection by the sampogrilate, as in the results of the animal test.

In addition, sarpogrelate can be administered in combination with Korean red ginseng extract, ginkgo biloba extract, resveratrol, vitis vinifera leaf dry extract, montelukast, etc., a composition for treating sensory neural deafness, which comprises sarpogrelate or a pharmaceutically acceptable salt thereof as an active ingredient, and a composition for treating sensory neuralgia, and a composition for treating ginsenosides, such as Korean red ginseng extract, ginkgo biloba extract, resveratrol, vitis vinifera leaf dry extract ), And montelukast. The synergistic effect of the combined administration of the combination preparation containing the active ingredient and at least one selected from the group consisting of montelukast, and montelukast can be obtained.

While the present invention has been particularly shown and described with reference to specific embodiments thereof, those skilled in the art will appreciate that such specific embodiments are merely preferred embodiments and that the scope of the present invention is not limited thereto will be. It is therefore intended that the scope of the invention be defined by the claims appended hereto and their equivalents.

Claims (17)

A pharmaceutical composition for the prevention or treatment of sensory neural deafness containing sarpogrelate or a pharmaceutically acceptable salt thereof as an active ingredient.
The pharmaceutical composition according to claim 1, further comprising at least one selected from the group consisting of Korean red ginseng extract, ginkgo biloba extract, resveratrol and vitis vinifera leaf dry extract. Composition.
The pharmaceutical composition according to claim 1, wherein the sensory neural deafness is a noise-induced hearing loss, a toxic hearing loss, a sudden hearing loss or an aging hearing loss.
2. The pharmaceutical composition according to claim 1, wherein the sensory neuron deafness is caused by damage of inner ear hair cells and surrounding tissues.
The pharmaceutical composition according to claim 1, which is characterized by increasing the suppression of apoptosis of auditory cells or the expression of antioxidant enzymes in auditory cells.
The composition of claim 1 wherein said pharmaceutically acceptable salt is selected from the group consisting of hydrochloride, sulfate, mesylate, malate, maleate, mesylate, besylate, hydrogen sulfate, oxalate, fumarate, tartrate, Wherein the salt is at least one salt selected from the group consisting of succinic acid salts, acetic acid salts, and phosphates.
The pharmaceutical composition according to claim 1, further comprising a pharmaceutically acceptable additive.
The pharmaceutical composition according to claim 7, wherein the additive is at least one selected from the group consisting of an excipient, a binder, a lubricant, a lubricant, a disintegrant, a sweetener, a flavor, a carrier and a mixture thereof.
A combination preparation for preventing or treating sensory-nerve-disorder deafness containing the composition of claim 2 as an active ingredient.
10. The combination preparation according to claim 9, which is a tablet, a foamable tablet, a granule, a powder, an injection, a pill or a capsule.
Consisting of Korean red ginseng extract, ginkgo leaf extract, resveratrol and vitis vinifera leaf dry extract containing sarpogrelate or a pharmaceutically acceptable salt thereof as an active ingredient A pharmaceutical composition for the prevention or treatment of sensory-nerve deafness for combination administration with at least one selected from the group.
12. The pharmaceutical composition according to claim 11, wherein said combination administration is carried out sequentially, simultaneously or in reverse order.
A food composition for preventing or ameliorating sensory neural deafness, comprising sarpogrelate or a pharmaceutically acceptable salt thereof as an active ingredient.
14. The food according to claim 13, further comprising at least one selected from the group consisting of Korean red ginseng extract, ginkgo biloba extract, resveratrol and vitis vinifera leaf dry extract Composition.
14. The food composition of claim 13, wherein the sensory neural deafness is a noise-induced hearing loss, a toxic hearing loss, a sudden hearing loss or an aging hearing loss.
14. The food composition according to claim 13, wherein the sensory neuron deafness is caused by damage of inner ear hair cells and surrounding tissues.
14. The food composition of claim 13, further comprising a pharmaceutically acceptable additive.

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